The long-range objective of this research effort is to determine the biochemical role of copper in iron, catecholamine, and energy metabolism, and to determine to what exten the pathological expression of copper deficiency is due to secondary iron deficiency. To accomplish these objectives and to solve certain enigmas that exist between genetic and environmental copper deficiency, female mice of the C3H/HeJ strain, heterozygous for the Brindled allele at the Mottled locus (Mobr/+) will be bred with normal males (Mo+/y). Homozygous male offspring (Mobr/y) have many features of copper deficiency (genetic copper deficiency). They will be compared with normal males (Mo+/y) which are copper-deficient because their dams (Mo+/+) are fed a copper-deficient diet (dietary copper deficiency). Offspring of copper-deficient rats will be used for some comparisons. Copper supplemented, control mice and rats will also be studied. Tissue copper and iron levels will be measured in young mice and rats to follow copper depletion and potential iron alterations. Hematological and enzymatic data will be collected from these same animals, including measurements of hemoglobin, hematocrit, ceruloplasmin, and ferroxidase. Cytochrome oxidase activity will be measured in erythropoeitic tissues. Mouse serum ceruloplasmin and ferroxidase will be partially purified and studied. These data in the mouse model will help to delineate the biochemical mechanisms of copper-dependent anemia and will help to resolve the enigma known to exist between copper-dependent anemia humans and infants with Menke's disease (genetic copper deficiency). Norepinephrine, dopamine, and ascorbic acid will be measured in mouse brain regions by HPLC with electrochemical detection before and after copper and iron supplementation. Regional measurements of copper and iron will be performed by graphite furnace atomic absorption spectroscopy. These data will be valuable in determining the role of copper and iron in catecholamine metabolism. Energy metabolism will be studied morphologically by examining tissue in the electron microscope, emphasizing mitochondria. The functional status of energy metabolism in brain will be assessed by measuring protein synthesis, in vitro, and following myelination. Metabolite analysis of nucleotides and glycolytic intermediates will enhance our understanding of copper-dependent mechanisms of aberrant energy metabolism.